FIG 8.
Cell surface expression of ORF3 protein and its mutants. Vero cells grown on coverslips were transfected with pORF3wt and ORF3 mutant constructs including AITF, YLAG, YLAA, ALAI, AKL, AATI, YATG, EAA, AAD, and ΔYLAI. After 36 h of incubation, the cells were fixed and processed for IFA under nonpermeabilization conditions by using P71-3 as primary antibody and Alexa Fluor 647-conjugated goat anti-rabbit IgG (red) as secondary antibody. Cells transfected with empty vector served as a negative control (mock). Cell nuclei were stained with DAPI (blue). Separate images showing surface ORF3 protein expression and nuclear staining are presented, as well as mergers of the two. Scale bar represents 15 μm.